DNA fingerprinting of Brucella abortus isolated from bovine brucellosis outbreaks by repetitive element sequence (rep)-PCR

  • Accepted : 2005.05.23
  • Published : 2005.06.30

Abstract

DNA fingerprint patterns of 8 Brucella reference strains and 15 B. abortus field isolates were characterized by repetitive element sequence-based PCR (rep-PCR) using BOX- and ERIC-primers in this study. AMOS PCR differentiated all Brucella field isolates from B. abortus RB51, a vaccine strain by producing a B. abortus-specific 498 bp band. Rep-PCR using BOX-primer produced 13 to 18 bands with sizes of between 230 and 3,300 bp, and discriminated Brucella strains to the species level except B. canis and B. suis. PCR products amplified with ERIC primers were, however, not appropriate for differentiating the Brucella isolates. DNA fingerprint patterns for all B. abortus field isolates were identical among them and were put on one cluster with B. abortus biovar 1 reference strain in the dendrogram, indicating they were highly clonal. These results suggested that rep-PCR using BOX primer might to be a useful tool for calculating genetic relatedness among the Brucella species and for the study of brucellosis epidemiology.

Keywords

References

  1. 국립수의과학검역원. 국립수의과학검역원 연보. pp. 79-90, 국립수의과학검역원, 안양 2003
  2. Allardet-Servent A, Bourg G, Ramuz M, Pages M, Bellis M, Roizes G. DNA polymorphism in strains of the genus Brucella. J Bacteriol 1988, 170, 4603-4607 https://doi.org/10.1128/jb.170.10.4603-4607.1988
  3. Alton GG, Jones LM, Pietz DE. Laboratory Techniques in Brucellosis. Monograph Series. World Health Organization, Geneva, Switzerland. 1975
  4. Bricker BJ, Ewalt AP, MacMillan AP, Foster G, Brew S. Molecular characterization of Brucella strains isolated from marine mammals. J Clin Microbiol 2000, 38, 1258-1262
  5. Bricker BJ, Halling SM. Differentiation of Brucella abortus bv. 1, 2, and 4, Brucella melitensis, Brucella ovis, and Brucella suis bv. 1 by PCR. J Clin Microbiol 1994, 32, 2660-2666
  6. Burr MD, Josephson KL, Pepper IL. An evaluation of ERIC PCR and AP PCR fingerprinting for discriminating Salmonella serotypes. Lett Appl Microbiol 1998, 27, 24-30 https://doi.org/10.1046/j.1472-765X.1998.00378.x
  7. Chung JS, Cho YJ, Park CK. Isolation and biotyping of Brucella abortus from dairy cattle in Kyungbuk area, Korea. Korean J Vet Res 1988, 28, 339-343
  8. Corbel MJ. Brucellosis: an overview. Emerg Infect Dis 1997, 3, 213-221 https://doi.org/10.3201/eid0302.970219
  9. Dombek PE, Johnson LK, Zimmerley ST. Use of repetitive DNA sequences and the PCR to discriminate Escherichia coli isolates from human and animal sources. Appl Environ Microbiol 2000, 66, 2572-2577 https://doi.org/10.1128/AEM.66.6.2572-2577.2000
  10. Ewalt DR, Bricker BJ. Validation of the abbreviated Brucella AMOS PCR as a rapid screening method for differentiation of Brucella abortus field strain isolates and the vaccine strains, 19 and RB51. J Clin Microbial 2000, 38, 3085-3086
  11. Fekete A, Bantle JA, Halling SM, Stich RW. Amplification fragment length polymorphism in Brucella strains by use of PCR with arbitrary primers. J Bacteriol 1992, 174, 7778-7783 https://doi.org/10.1128/jb.174.23.7778-7783.1992
  12. Ficht TA, Husseinen HS, Derr J, Bearden SW. Species-specific sequences at the omp2 locus of Brucella type strains. Int J Syst Bacteriol 1996, 46, 329-331 https://doi.org/10.1099/00207713-46-1-329
  13. Jensen AE, Cheville NF, Thoen CO, MacMillan AP, Miller WG. Genomic fingerprinting and development of a dendrogram for Brucella spp. Isolated from seals, porpoises, and dolphins. J Vet Diagn Invest 1999, 11, 152-157 https://doi.org/10.1177/104063879901100208
  14. Jersek B, Gilot P, Gubina M, Klun N, Mehle J, Tcherneva E, Rijpens N, Herman L. Typing of Listeria monocytogenes strains by repetitive element sequence-based PCR. J Clin Microbiol 1999, 37, 103-109
  15. Mercier E, Jumas-Bilak E, Allardet-Servent A, O'Callaghan D, Ramuz M. Polymorphism in Brucella strains detected by studying distribution of two short repetitive DNA elements. J Clin Microbiol 1996, 34, 1299-1302
  16. Michaux-Charachon S, Bourg G, Jumas-Bilak E, Guigue-Talat P, Allardet-Servent A, O'Callaghan D, Ramuz M. Genome structure and phylogeny in the genus Brucella. J Bacteriol 1997, 179, 3244-3249 https://doi.org/10.1128/jb.179.10.3244-3249.1997
  17. O'Hara MJ, Collins DM, de Lisle GW. Restriction endonuclease analysis of Brucella ovis and other Brucella species. Vet Microbiol 1985, 10, 425-429 https://doi.org/10.1016/0378-1135(85)90024-0
  18. Olsen JE, Skov MN, Threlfall EJ, Brown DJ. Clonal lines of Salmonella enterica serotype Enteritidis documented by IS200-, ribo-, and pulsed field gel electrophoresis and RFLP typing. J Med Micribiol 1994, 40, 15-22 https://doi.org/10.1099/00222615-40-1-15
  19. Rijpens NJ, Jannes G, Van Asbroeck M, Rossau R, Herman LM. Direct detection of Brucella spp. In raw milk by PCR and reverse hybridization with 16S-23S rRNA spacer probes. Appl Environ Microbiol 1996, 62, 1683-1688
  20. Tcherneva E, Rijpens N, Jersek B, Herman LMF. Differentiation of Brucella species by random amplified polymorphic DNA analysis. J Appl Microbiol 2000, 88, 69-80 https://doi.org/10.1046/j.1365-2672.2000.00945.x
  21. Tcherneva E, Rijpens N, Naydensky C, Herman L. Repetitive element sequence based on polymerase chain reaction for typing of Brucella strains. Vet Microbiol 1996, 51, 169-178 https://doi.org/10.1016/0378-1135(96)00036-3
  22. Thong KL, Ngeow YF, Altwegg M, Navaratnam P, Pang T. Molecular analysis of Salmonella enteritidis by pulsed-field gel electrophoresis and ribotyping. J Clin Microbiol 1995, 33, 1070-1074
  23. Verger JM, Grimont F, Grimont PAD, Grayon M. Brucella, monospecific genus as shown by deoxyribonucleic acid hybridization. Int J Syst Bacteriol 1985, 35, 292-295 https://doi.org/10.1099/00207713-35-3-292
  24. Versalovic J, Koeuth T, Lupski JR. Distribution of repetitive DNA sequences in eubacteria and application to fingerprinting of bacterial genomes. Nucelic Acids Res 1991, 19, 6823-6831 https://doi.org/10.1093/nar/19.24.6823
  25. Vizcaino N, Verger JM, Grayon M, Zygmunt MS, Cloeckaert A. DNA polymorphism at the Omp-31 locus of Brucella spp. Evidence for a large deletion in B. abortus, and other species-specific markers. Microbiology 1997, 143, 2913-2921 https://doi.org/10.1099/00221287-143-9-2913
  26. Weigel RM, Qiao B, Teferedegne B, Suh DK, Barber DA, Isaacson RE, White BA. Comparison of pulsed field gel electrophoresis and repetitive sequence polymerase chain reaction as genotyping methods for detection of genetic diversity and inferring transmission of Salmonella. Vet Microbiol 2004, 100, 205-17 https://doi.org/10.1016/j.vetmic.2004.02.009
  27. Welsh J, McClelland M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 1990, 18, 7213-7218 https://doi.org/10.1093/nar/18.24.7213
  28. Williams E. Brucellosis in humans-its diagnosis and treatment. APMIS Suppl 1988, 3, 21-25